KR20180055945A - Composition for Improving hepatic function Including Extract of Salvia Militorhiza Bunge and Method for Preparation of the Same - Google Patents

Abstract

The present invention relates to a composition for improving a hepatic function including a root extract of Salvia miltiorrhiza Bunge as an active ingredient, wherein the root extract of Salvia miltiorrhiza Bunge contains 20% or more salvianolic acid B. The composition additionally includes an aloe extract and a noni extract in addition to the root extract of Salvia miltiorrhiza Bunge.

Description

TECHNICAL FIELD The present invention relates to a composition containing a rosemary extract for improving liver function and a method for preparing the same.

The present invention relates to a composition containing a root extract for improving liver function and a method for producing the same.

The liver, which is an indispensable organ for life such as metabolism, detoxification and storage of influenza, has more than 500 functions and maintains the normal function of the human body by the action of various enzymes of 2000 kinds or more. Therefore, Liver function improvement has a close relationship with human health, and researches on it have been actively conducted.

Specifically, symptoms that occur when a liver function disorder occurs include, for example, alcoholic fatty liver in which a large amount of neutral fat accumulates in the liver due to frequent alcohol consumption, alcoholic hepatitis causing hepatocyte necrosis or denaturation, and hepatocyte necrosis Alcoholic hepatopathy such as alcoholic hepatic encephalopathy, which is increased only in the skin; Fatty liver in which a large amount of fat is accumulated in hepatocytes by obesity or metabolic diseases; Cirrhosis in which hepatic cells lose their elasticity due to collagen, a fibrous component produced when the destroyed hepatocytes are regenerated, and become stiff and hardened; Hepatitis, which is an inflammation of hepatocytes and liver tissues due to viruses, alcohols, drugs and autoimmunity; And liver cancer, which is a malignant tumor originating from hepatocytes.

In addition, when a problem occurs in the liver function as described above, symptoms such as stain, pigmentation, aging promotion, jaundice, food insufficiency, and weight loss occur, and it is important to treat early symptoms when the initial symptoms occur.

Salvianolic acid B is widely known as a substance useful for such liver function. Especially, Dansam has a lot of pharmacological components such as salvianone and tanshinone, Is known to be effective.

It is the root of salvia miliorrhiza bunge , which is the standard of herbal medicine. The average content of Salvianolic acid B (Salvianolic acid B) is 4.1% B content is less than 5% on average. Therefore, studies for obtaining a composition containing salvianoic acid B in a high content in order to obtain a high liver function improving effect are continuing.

Specifically, the selected root is roasted and crushed, treated with an enzyme to increase the recovery rate, and then extracted using a 95% alcohol. Thereafter, filtration and concentration processes are performed through filtering.

However, since the content of the active ingredient, salvianoic acid B is only a small amount, there is a problem that a large amount of the extract is required in order to exhibit a desired effect.

Therefore, in order to improve the liver function improving effect, there is a high necessity to develop a dansam root extract having a high active ingredient content and its preparation method.

SUMMARY OF THE INVENTION It is an object of the present invention to solve the above-mentioned problems of the prior art and the technical problems required from the past.

The inventors of the present application have conducted intensive studies and various experiments and, as will be described later, a composition for improving liver function comprising Dansamp roots extract as an active ingredient, wherein the content of salvianoic acid B is 20% or more The present invention has been accomplished on the basis of the discovery that the addition of Aloe extract and Noni extract in addition to Dansamp root extract has a high liver function improving effect.

In order to achieve the above object, the composition for improving liver function according to the present invention comprises a dansam root extract containing salvianoic acid B at a high concentration. Specifically, the composition for improving liver function comprises Salvianone B The extract contains Dansamp roots extract, Aloe extract, and Noni extract containing 20% or more of the content.

In general, the content of salvianolic acid B in salvianolic acid B is 4.1% or more on average in dried products. As a method of extracting salvianoic acid B, conventionally, raw materials of radish sprouts are selected, raw materials for washing and grinding are selected The enzymatic treatment of cellulose enzymes at 40 ° C for 6 hours, the addition of 10% of the weight of dry matter to the extracts for 12 hours, the extraction of the supernatant of Dansamp roots extract, The process of filtration is to concentrate the recovered solution to 30 brix or more using a reduced pressure concentrator, freeze-drying the concentrated sample at -40 ° C, and pulverizing and lyophilizing the freeze- And a selection process.

As a result of the extraction according to the above method, the content of the indicator material is analyzed as shown in Table 1 below.

Analysis item Measurement result (%) Salvianaconic acid B 8.18 Tanshinon II A 0.014%

As shown in Table 1, the content of salvianoic acid B was 8.18%, which corresponds to about 584 times of 0.014% of Tanshinone II A, which is effective for the prevention and treatment of metabolic syndrome. The amount of salvianoic acid B extracted is contained at a high concentration compared with the content of other indicator materials.

Furthermore, the inventors of the present invention have conducted an experiment for extracting salvianoic acid B at a higher concentration using dansam root, and as a result, developed a method for obtaining dansam root extract containing at least 20% of salvianoic acid B .

Specifically, the method comprises the steps of (a) pulverizing washed root, (b) drying the pulverized root, and (c) extracting and drying the root extract, And (d) drying the concentrated extract.

Specifically, the extraction process of step (b) may be performed at 40 ° C to 90 ° C for 5 hours to 20 hours, and may be selectively performed in an appropriate range depending on the type of the solvent used.

In addition, in order to obtain a dansam root extract containing a high concentration of salvianoic acid B, the kind of extraction solvent is very important. The extraction solvent may be water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof. Lower alcohols having 1 to 4 carbon atoms, or a mixed solvent thereof, and more specifically 70 to 90% alcohol can be used.

The step (d) may be any one of natural drying, vacuum drying, air drying, air drying, hot air drying, flow drying, spray drying, freeze drying, infrared drying and high frequency drying. In order to prevent the active ingredient from being destroyed , The freeze-drying method is preferably used.

Meanwhile, the method may further include a step (e) of pulverizing and pulverizing the powder. The dried powder may be pulverized and pulverized, and the pulverized powder may be pulverized to 80mesh or less.

As an example of a specific method for producing dansam root extract, a raw material selection process for screening, washing and pulverizing the raw materials of dansam root, drying the pulverized dansam root, adding 10 times 80% 12 hr extraction, recovery of the supernatant of Dansamp roots extract, filtration using a 1 μm filter, concentration of the recovered solution to 30 brix or more using a reduced pressure concentrator, and freezing of the concentrated sample at 40 ° C And a drying process.

As a result of the extraction according to the above method, the content of the indicator material is analyzed as shown in Table 2 below.

Analysis item Measurement result (%) Process yield 15.1 Salvianaconic acid B 20 or more

Referring to Table 2, it can be seen that high concentration of salvianoic acid B can be obtained as well as high yield of the process as a result of eliminating the enzyme treatment process and different types of extraction solvent.

On the other hand, in order to develop a raw material having an excellent effect on improvement of liver function based on the raw material of dansam root extract, it is possible to consider mixing with various raw materials in the dansam root extract prepared by the production method of dansam root extract. As such a sub ingredient, a material exhibiting a synergistic effect in a state of excellent liver function improving effect and mixed with a root extract is suitable, and noni fruit and aloe avoresense leaves can be used as materials securing the safety of raw materials.

In order to extract the active ingredients contained in the noni fruit and aloe avocence leaf, each of the raw materials was extracted using a 70% alcohol for 12 hours at 65 ° C and filtered using a 50 μm filter. Is carried out in the same manner as the extraction process of the active ingredient of the high-concentration root extract.

The mixed extract of Panax notoginseng extract may be prepared by mixing 10 to 30% by weight of Panax ginseng extract, 45 to 45% by weight of aloe extract 45 And more preferably 20% to 30% by weight of noni extract, 15% to 25%, 50% to 60% of aloe extract and 22% to 27% of noni extract Lt; / RTI >

The salvianoic acid B, which is an active ingredient of the dansam root extract, may be represented by the following structural formula (1), and the active ingredient of the aloe extract may be represented by the structural formula (2) as Barbaloin, Scopoletin, an active ingredient of the noni extract, can be represented by the following structural formula (3).

(1)

(One)

(2)

(2)

(3)

(3)

Meanwhile, the inventors of the present invention have experimented with various factors related to the liver function improving effect through various methods using the above-mentioned Dansampo extract, and the details thereof are as follows.

In vivo liver function improvement experiment of Dansam mixed extract

The high fat diet using rodent induction fatty liver model is considered to be suitable for the nonalcoholic fatty liver model because it induces fatty liver according to the metabolic syndrome induced by diet. Therefore, in this study, we aimed to observe changes in liver function and hepatic function - related index after ingestion of Panax ginseng extract in high fat diet - induced obese rats.

As a control substance, a positive control substance (200 mg / kg of daily dose) was fed with a high-fat diet-fed control and a silymarin-fed high fat diet, and 1%, 2% High - fat diets containing 3% Scotch - Blend mixed extract were used.

Dietary Obesity Induction (DIO)

Three weeks old male C57BL / 6 mice were obtained and fed with commercial mouse feed and negative water for 1 week. After adjusting to the breeding environment, high fat diets fed with 45% calories as fat (D12451, Research Diet , USA) for 4 weeks to induce obesity. After 4 weeks, test group was formed according to body weight. The control group was fed with only high fat diet and the positive control group was fed with 200 mg / kg of daily dose of silymarin mixed with high fat diet and fed for 8 weeks. In the test group, 1%, 2% % And 3%, and fed for 8 weeks.

Changes in blood glucose and lipid composition

Total cholesterol, HDL-cholesterol and LDL-cholesterol in the lipid components were measured by collecting blood from the abdominal vein under fasting condition for 12 hours. Serum total cholesterol, HDL-cholesterol, and LDL-cholesterol levels were decreased in the control group and in the test group fed the silymarin-fed control group and the monosan mixed extract group, respectively. In the test group, , Respectively.

FIG. 1 is a graph showing changes in blood LDL-cholesterol concentration in experimental animals according to high-fat diets and sample intake for 8 weeks. The concentration of LDL-cholesterol was lower in the control group than in the control group, but the difference was not statistically significant. However, the test group fed the mixed extract of Dansamp showed a statistically significant decrease (p <0.05) There was no significant difference between the mixed extracts.

Changes in liver function and renal function indices

ALT (alanine transaminase) as an index of liver function was measured and compared with the serum of experimental animals. The results are shown in Table 1 and FIG. 2 below. As shown in Table 1 and FIG. 2, serum ALT levels measured as a measure of liver damage did not show a significant difference in the positive control group fed with silymarin against the negative control group, but the feeding amount of the extract group And 2% and 3%, respectively, of the mixed extract of Panax notoginsenoside showed a statistically significant decrease in the negative control group and the positive control group. (p < 0.05)

Groups ALT (IU / L) Negative Control (Control) 66.1 ± 43.9 ^a The positive control (silymarin) 66.7 ± 32.1 ^a 1% water extract 49.1 ± 23.3 ^ab 2% water extract 37.4 ± 23.6 ^b 3% water extract 26.9 ± 7.2 ^b

1) ^{a, ab, b} , different superscript means statistical significance: p <0.05

Changes in serum adipokine-based index factors

Adipokine-based index factors such as adiponectin, insulin, leptin, and resistin were measured by immunochemical methods as indicators of liver function damage due to high fat diet from serum of experimental animals after feeding for 8 weeks with high fat diet and experimental diet The results are shown in Table 4 and FIG. 3 below.

Groups Adiponectin 1)
(ng / mg) Insulin
(ng / ml) Leptin
(ng / ml) Resistin
(ngl / ml) Negative Control (Control) 5.2 ± 12.1 4.1 ± 1.7 49.5 ± 16.8 ^{a2 )} 5.0 ± 1.0 The positive control (silymarin) 6.4 ± 8.5 1.9 ± 1.2 18.9 ± 4.4 ^b 3.2 ± 1.4 1% water extract 8.1 ± 7.2 2.3 ± 2.1 21.9 ± 9.2 ^b 4.2 ± 1.3 2% water extract 6.3 ± 9.4 2.9 ± 1.8 23.9 ± 10.6 ^b 4.1 ± 1.3 3% water extract 6.7 ± 6.0 2.1 ± 1.9 21.9 ± 11.5 ^b 4.3 ± 1.8

1) Adiopunctin (ng / mg) = [serum adiponectin contents (μg / ml)] / [adipose tissue weight (fat mass, mg)]

2) ^{a, ab, b} , different superscript means statistical significance: p <0.05

Adiponectin, insulin, leptin, and resistin were increased in adiponectin concentration and insulin, leptin and resistin concentrations were decreased in control group and silkworm - treated group. .

3, leptin concentration was significantly lower in the control group than in the control group and in the test group fed the mixed extract (p <0.05) As shown in FIG. 4, the ratio of leptin to adiponectin (L / A ratio), which is observed as a major factor of the present study, is comparatively evaluated. As shown in FIG. 4, the negative control group and the control group fed with 1% (P <0.05), and the experimental group fed 2% and 3% of the mixed extracts showed a significantly lower tendency.

Experimental Mechanism of In vitro Liver Function Enhancement of Dansam Mixed Extract

In order to investigate the mechanism of action of hepatic ginseng extract on hepatic function improvement in vitro by HepG2 cell line, hepatic cell line, AMPK, ACC, and Nrf2, and antioxidant activity.

Evaluation of AMPK activity control ability

AMPK and ACC (acetyl-CoA) were added to the HepG2 cell line for 6 hours at a concentration of 30 μg / ml in the presence of aloe extract (eA), noni extract (eN) carboxylase activity was evaluated. The results are as follows.

<Evaluation of the effect of Dansam mixed extract on AMPK activity in HepG2 cell line>

All of the aloe extract, noni extract, ginseng extract and ginseng mixed extract increased the activity of AMPK and inhibited the activity of ACC on the control group. In particular, I could.

On the other hand, among the single extracts, the extracts of A. ginseng showed higher AMPK activity and ACC inhibitory activity on aloe and noni extracts, and the control ability of noni extract was lowest.

In addition, AMPK activity and ACC inhibition activity were found to be maximal up to 6 hours after treatment with 100 μg / ml of HepG2 cell line.

<Evaluation of effect of AMPK activity of HepG2 cell line over time>

The concentration of AMPK in the concentration of 0, 10, 30, and 100 μg / ml in the HepG2 cell line and the concentration of AMPK was 100 μg / ml, .

<Evaluation of the effect of Dansam mixed extract on AMPK activity in HepG2 cell line>

The expression levels of sirt1, carnitine palmitoyltransferase (CPT) and ACC (acetyl-CoA carboxylase) involved in AMPK activity were measured by real-time qPCR in a HepG2 cell line for 24 h at a concentration of 100 μg / As shown in FIG. 5, the mixed extract of Dansamp showed activation of sirt1 and significantly inhibited the production of CPT and ACC.

The expression level of sirt1, which is involved in AMPK activity, was measured by real time after 10, 30, and 100 μg / ml of HepG2 cell line was treated with real -time &lt; / RTI &gt; qPCR. The results are shown in Fig. Referring to FIG. 6, the extracts of Dansamp mixture increased the expression rate of sirt1 in a dose-dependent manner and showed the highest activity at a concentration of 100 μg / ml.

Evaluation of Nrf2 activity regulation ability

In order to evaluate the ability of Nrf2 (Nf-E2-related factor-2) to regulate the activity of aloe extract, ginseng extract, noni extract and radix extract, the expression of HO-1 (heme oxygenase-1) and the results are shown in FIG.

The gene expression levels of HO-1 and NQO-1 (NAD (P) H: quinone oxidoreductase) were measured at 3, 6, 12 and 24 hours after treatment with HepG2 cell line at a concentration of 100 μg / The activity of HO-1 was increased in all the samples. The activity of HO-1 was significantly increased up to 12 hours, and NQO-1 6 hours. &Lt; / RTI &gt;

The gene expression levels of HO-1 and NQO-1 (NAD (P) H: quinone oxidoreductase) were measured by real-time qPCR using HepG2 cell lines at 10, 30 and 100 μg / The results are shown in FIG. 8. As shown in FIG. 8, the expression level of HO-1 was significantly increased in the concentration-dependent manner after the treatment with the extracts of Alnia, Radix, and Noni extracts .

The expression levels of Nrf2 and PARP (poly (ADP-ribose) polymerase) of nucla- rer and cytosol were measured after treatment with 10, 30 and 100 μg / As a result of PCR, Nrf2 and PARP increased in a concentration dependent manner as shown in the figure below.

Assessment of antioxidant and anti-inflammatory effects

The results obtained by measuring the DPPH radical scavenging activity after treatment with the extracts of Dansamp mixture at a concentration of 10, 10 and 100 μg / ml in the HepG2 cell line are shown in FIG. 9, and the amounts of reactive oxygen species (ROS) The results are shown in FIG. 10, and the amount of reduction of GSH (glutachione) according to the concentration of the raw ginseng mixed extract is shown in FIG.

The radical scavenging ability and reduced GSH were increased and the ROS production in cells was decreased depending on the concentration.

The concentrations of NO (nitric oxide) and IL-6 produced by LPS-induced inflammation in RAW264.7 cell line treated with 10, 30, and 100 μg / Table 5 shows that NO production and IL-6 production are significantly decreased in a concentration-dependent manner.

Group NO (uM) IL-6 (pg / ml) Con 11.43 + - 0.24 2.29 ± 0.56 LPS 27.37 ± 1.31 116.41 + - 23.55 10 27.85 + - 0.62 102.10 ± 17.17 30 20.89 ± 0.48 91.22 + - 18.14 100 13.38 ± 0.24 62.10 ± 9.68

In addition, the RAW264.7 cell lines were treated with 10, 30, and 100 μg / ml of extracts of Panax ginseng extracts. The concentrations of iNOS (inducible nitric oxide synthase) and COX-2 (cyclooxygenase- 2) were measured, and the expression levels of iNOS and COX-2 were significantly decreased in a concentration-dependent manner.

As a result of the above experiment, the mechanism of liver function improvement of the mixed extract of Panax notoginseng using the in vitro evaluation model was confirmed, and it was found that the mixed extract of Panax ginseng was treated with a single raw material such as aloe extract, The activity of AMPK was significantly higher than that of ATPK, but the activity of AMPK was significantly inhibited in sirt1 and CPT.

The activity of HO-1 was significantly influenced by the activity of Nrf2, which is involved in the detoxifying function of liver, than that of single raw materials such as Aloe extract, Root extract and Noni juice. . The expression of HO-1 and NQO-1 was significantly increased in a concentration-dependent manner and the expression of Nrf2 and PARP in the nucleus was also significantly increased.

In the evaluation of antioxidant activity as described above, the mixed extract of Dansamp showed increased scavenging activity and GSH activity of DPPH radicals and ROS in cells and inhibited the production of NO and IL-6, which are factors related to inflammation, and inhibited iNOS and COX -2. &Lt; / RTI &gt; In conclusion, according to the results of this study, it was confirmed that the mixed extract of Panax ginsenosides protects and improves the liver function by controlling Nrf2 and antioxidant and antiinflammatory effects which are involved in the hepatotoxicity of AMPK and liver, do.

As described above, the composition for improving liver function according to the present invention comprises the extract of Dansamp roots, aloe extract, and noni extract, which contains a high content of salvianoic acid B, an effective ingredient for improving liver function, There is an effect of reducing the total cholesterol concentration in the blood, reducing the index of liver function damage, and improving liver function.

In addition, the extract of Dansamp roots containing a high content of salvianoic acid B, which is an active ingredient for improving liver function, can be produced, and it can be developed as a health functional food for improving liver function.

FIG. 1 is a graph showing changes in blood LDL-cholesterol concentration in experimental animals according to high fat diets and sample intake for 8 weeks;
FIG. 2 is a graph showing changes in liver damage index ALT of experimental animals according to high-fat diets and sample intake for 8 weeks; FIG.
FIG. 3 is a graph showing changes in blood letin of experimental animals according to high-fat diets and sample intake for 8 weeks; FIG.
FIG. 4 is a graph showing changes in the leptin / adiponectin ratio of experimental animals with high fat diets and sample intake for 8 weeks;
FIG. 5 is a graph showing the effect of the extract of Panax ginseng on the expression levels of sirt1, carnitine palmitoyltrnasferase and acetyl-CoA carboxylase in the HepG2 cell line;
FIG. 6 is a graph showing the effect of the treatment of concentration of Dansampo extract on the expression level of sirt1 in the HepG2 cell line;
FIG. 7 shows the effect of concentration and time of treatment on the expression level of HO-1 in the HepG2 cell line of aloe extract, ginseng extract, noni extract and mixed ginseng extract;
FIG. 8 is a graph showing the effect of treatment of the concentration of RDA extract on the expression level of HepG2 cell line HO-1 and NQO-1;
FIG. 9 is a graph showing DPPH radical scavenging ability according to the treatment of the concentration of Panax ginseng mixed extract;
FIG. 10 is a graph showing the evaluation of reactive oxygen species (ROS) production in cells of the HepG2 cell line according to the treatment of the concentration of the extracts of R. radix;
FIG. 11 is a graph showing the effect of concentration-dependent treatment of Rawan extract on the reduction of GSH (glutachione) in the HepG2 cell line;
FIG. 12 is a graph showing changes in survival rate of experimental animals after administration of LPS / D-galN peritoneal cavity through hepatic toxicity;
FIG. 13 is a graph showing changes in the expression rate of HO-1 in experimental animals after administration of LPS / D-galN peritoneal toxin;
14 is a photograph (a) of a H & E staining tissue slide (× 400) of hepatic toxicity induced by LPS / D-galN intraperitoneal administration of experimental liver tissue;
FIG. 15 is a photograph (b) of a TUNEL DAB stained tissue slice (× 400) of hepatic toxicity-induced experimental liver tissue through LPS / D-galN peritoneal administration; And
FIG. 16 is a graph showing absorption of salvianolic acid and evaluation of pharmacokinetics of metabolism until 24 hours after ingestion of the extracts of Panax ginseng and Panax ginseng. FIG.

Hereinafter, the contents of the present invention will be described in detail by way of examples of the present invention, but the present invention is not limited by the scope of the present invention.

Experimental Example 1 In vivo liver function improvement experiment

To evaluate the effects of treatment with LPS (lipopolysaccharide) and galactosamine on hepatic toxicity in male rats, 5-week-old male ICR mice were classified into each group. The control and negative controls were fed only AIN76A standard diet and the positive control group was fed with AIN76A feed at a dose of 200 mg / kg of daily intake of silymarin and fed for 2 weeks.

As a test material, D-galactosamine 700 mg / kg and LPS 10 μg / kg were administered intraperitoneally (intraperitoneally) to the mice after 2 weeks of feeding the mixture of monascus mixed extracts containing 1%, 2% and 3% IP) to induce hepatotoxicity.

Measurement of HO-1 expression in liver tissue

1) To evaluate the expression of HO-1 (heme oxygenase-1) in liver tissues, RNA was extracted from liver tissue, and then cDNA was synthesized using Maxime RT premix (iNtRON) with 1 μg of RNA. TOPreal qPCR 2X PreMIX real-time PCR was performed using the enzymes of the present invention under the following conditions. 5'-GCGTGCAAGGGATGATTTCC-3 ', mouse HO-1 forward primer: 5'-CGCAACAAGCAGAACC-CA-3', mouse HO-1 reverse primer: 5'-GCGTGCAAGGGATGATTTCC-3 ' After 1 minute of amplification step, total 40 amplifications were confirmed.

Histopathological examination

1) Fixation: Frozen section slices were prepared by freezing the soy sauce for each individual and then performing the feezing section using cryotomb.

2) H & E staining: The tissue slides were stained with hematoxylin solution for 10 minutes, washed with tab water, neutralized with ammonia, stained with Eosin for 5 minutes, decolorized with xylene, fixed on canada balsam, and observed with optical microscope.

3) Tunel assay: The liver tissue was immobilized in 4% formaldehyde solution, treated with proteinase K, stained with TdT and DAB, dehydrated in ethanol and xylene, and observed with an optical microscope.

Observation of survival rate of experimental animals

The survival rate of experimental animals was evaluated by inducing hepatotoxicity by intraperitoneal administration of LPS and galactosamine to experimental animals fed AIN76A feed, silymarin, and mixed extracts of Panax ginseng at various concentrations. Is shown in Fig.

In the control group, the survival rate was significantly increased up to 30 hours after the administration of LPS and galactosamine in the test group in which 1, 2 and 3% of the mixed extracts were fed for 2 weeks.

Changes in blood cytokines, ALT and AST

The results of measurement of TNF-α, IL-6, IL-1β and ALT and AST as indicators of blood hepatotoxicity as inflammatory cytokines after inducing hepatotoxicity by LPS and galactosamine in experimental animals are shown in Table 6 below.

TNF-a (pg / ml) IL-6 (pg / ml) IL-1? (Pg / ml) ALT (U / L) AST (U / L) Control 44.5 ± 10.8 25.0 ± 1.4 9.2 ± 1.2 35.7 ± 2.1 51.2 ± 7.4 LPS / GalN 0% 1296.2 ± 267.4 811.2 + - 76.7 4765.2 + - 812.6 6128.6 + - 742.8 51.7 ± 9.3 LPS / GalN One% 527.9 ± 153.8 295.2 ± 81.2 780.1 + - 254.7 1048.3 ± 226.8 31.0 ± 7.1 LPS / GalN 2% 341.0 + - 83.2 121.5 ± 43.6 484.3 ± 95.5 515.7 ± 73.4 8.7 ± 1.5 LPS / GalN 3% 286.3 ± 196.4 264.9 ± 125.6 527 ± 76.4 784.1 ± 58.5 7.2 ± 0.5 LPS / GalN Milk Th 1145.5 ± 31.91 801.3 + - 146.8 5213.9 ± 1273.4 5896.3 + - 827.4 46.8 ± 15.3

As shown in Table 6, the levels of TNF-α, IL-6 and IL-1β in the experimental animals fed the mixed extract of Panax ginseng were significantly decreased in the negative control group, 6 and IL-1β, respectively, showed a tendency to decrease in a dose-dependent manner, whereas IL-6 showed a lower tendency in the 2% intake group. In addition, the results of ALT and AST indicate that serum liver index is significantly reduced.

Changes in expression of HO-1 in liver tissues

The expression level of HO-1 in liver tissues of experimental animals is shown in FIG. 13. The expression of HO-1 was increased according to the intake of the mixed extract of Dansamp. Especially, the highest increase was observed in the 2% intake group appear.

Liver tissue histopathological examination

FIG. 14 shows the results of H & E staining of the liver tissues of the experimental animals. The negative control group with hepatotoxicity was severely damaged in liver tissues, and the experimental animals fed with silymarin showed some recovery On the other hand, it was observed that the liver tissues were significantly improved in the experimental animals fed the mixed extract of Dansamp.

On the other hand, the results of evaluation of liver tissue apoptosis by TUNEL DAB staining on the same slide are shown in FIG. 15. As shown in FIG. 15, the positive control group treated with silymarin seems to be somewhat restored to the negative control group, The experimental animals showed significant improvement in liver tissue, and the 3% intake group was improved to the normal control level.

In vivo liver function improvement experiment, mortality was observed up to 30 hours after induction of hepatotoxicity by LPS and galactosamine in experimental animals and survival rate was significantly increased in experimental animals fed monosynthesized mixed extract to negative control group. AST and ALT, which are the indicators of blood liver function, showed that the experimental animals fed the mixed extract of Panax ginseng showed decreased liver function with a decrease in the intake dose. TNF-α, IL-6 and IL-1β were also decreased by ingestion of the mixed extracts, while IL-6 was not significantly different in the 3% dose-receiving experimental animals.

The expression rate of HO-1 (heme oxygenase-1), which plays a major role in the regulation of Nrf2 in the liver, was significantly increased according to the intake of the mixed extract of Panax ginseng and H & E staining . As a result, it was shown that the damage of liver tissue of experimental animals fed with the mixed extract of Panax ginseng was significantly reduced in the negative control group. In addition, TUNEL DAB staining showed that liver tissue necrosis was significantly reduced in the negative control group compared with the control group.

Therefore, it can be seen from the results of the above experiment that the mixed extract of Danamus improves liver function by regulating Nrf2-related factors and controlling secretion of inflammatory cytokines against hepatotoxicity caused by LPS and galactosamine.

EXPERIMENTAL EXAMPLE 2 In vivo PK Mechanism Experiment of Dansampan Extract Extract

In order to evaluate pharmacokinetics related to the in vivo absorption metabolism of the indicator substance against oral ingestion of the mixed extract of Danams, 6-week-old male SD rats were obtained as experimental animals and fed commercial diets in a 1-week breeding environment After adaptation, the fasting was maintained for 12 hours before feeding the experimental material. The extracts of Dansamp extract and Panax ginseng mixture were suspended in sterilized distilled water and fed orally by oral gavage at a concentration of 500 mg / kg of extracts of Panax ginseng CA Meyer. From 0 to 24 hours after feeding, Blood samples were collected in a 2K EDTA-treated test tube, centrifuged at 1,800rpm for 15 minutes, and plasma was separated and analyzed.

FIG. 16 shows the results of analyzing the change of salvianolic acid B from blood plasma obtained after oral administration of 500 mg / kg of water extract of DS and DS.

16, the concentration of salvianolic acid B, the indicator substance of the extract of Panax ginseng, showed the highest peak at 0.33 hour and the concentration of the indicator substance of the Panax ginseng extract was higher in Panax ginseng extract. In addition, Salvianolic acid B showed a tendency to decrease from peak time to 24 hours, and the extracts of Mandan extract were almost excreted at 24 hours. However, the extracts of Mandan extract retained its concentration after 4 hours even at 24 hours.

Thus, the concentration of salvianolic acid B in blood was found to be maintained at the peak level and during the metabolic process up to 24 hours in the experimental animals fed with single - Therefore, it can be seen that the formulation of the mixed extract of Panax ginseng significantly increases the bioavailability of the indicator material with respect to the single raw material.

It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims.

Claims (9)

A composition for improving liver function comprising roots extract of Danshen as an active ingredient, wherein the Dansamp root extract contains Salvianolic acid B (B) in an amount of 20% or more, and the Dansamp roots extract A composition for improving liver function characterized by being a mixture of extracts of Aloe (Aloe) and noni (Noni). [Claim 2] The method according to claim 1, wherein the water-extractant mixture comprises 10 to 30% by weight of the extract of Dansamp methods, 45 to 65% by weight of aloe extract and 20 to 30% &Lt; / RTI &gt; The method according to claim 1, wherein the RDA extract promotes the secretion of adiponectin and reduces the ratio of insulin, Leptin, Resistin, and lectin to adiponectin (L / A ratio) Wherein the composition is a composition for improving liver function. 10. A method for producing the root extract of Danshams according to claim 1,
(a) pulverizing the washed root of dansam;
(b) a step of drying and then extracting the ground radish root;
(c) a step of filtering and concentrating the root extract of Dansamp; And
(d) drying the concentrated extract;
Wherein the method comprises the steps of: 5. The method according to claim 4, wherein the step (b) is carried out at 40 to 90 ° C for 5 to 20 hours. [5] The method according to claim 4, wherein the step (b) comprises using water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof. The method according to claim 4, wherein the step (d) comprises any one of natural drying, vacuum drying, air drying, air drying, hot air drying, flow drying, spray drying, freeze drying, infrared drying and high frequency drying A method for producing a root extract. [5] The method according to claim 4, wherein the preparation method further comprises a step (e) of pulverizing and pulverizing. A method for preparing a mixed extract of Dansamp roots, which comprises mixing a nonylpiperidine extract and an aloe extract with the root extract of Dansamp root prepared by the method of manufacturing a root extract according to claim 4.

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